What question did this study set out to answer?

To establish an efficient CRISPR-Cas9 gene editing platform to generate recombinant African swine fever viruses and overcome limitations of traditional homologous recombination.

February 12, 2018Scientific ReportsOpen Access

CRISPR-Cas9, a tool to efficiently increase the development of recombinant African swine fever viruses

Q: What is the clinical evidence from this study?

Study design: Other. Population: African swine fever virus (ASFV). Intervention: CRISPR-Cas9 gene editing system vs. Traditional homologous recombination. Primary outcome: Frequency of recombination (ratio of recombinant to parental viruses).

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Key result

CRISPR-Cas9 increased the efficiency of developing recombinant African swine fever viruses, achieving a recombination frequency of 1 in 122 to 178 compared to <1 in 1,000,000 with traditional methods.

Population

Swine macrophages infected with highly virulent field strain Georgia07 of African swine fever virus (ASFV)

Comparison

CRISPR-Cas9 gene editing system to delete the… vs Traditional homologous recombination

Design

Preclinical

Authors

MBManuel V. BorcaNational Agricultural Statistics Service LHLauren G. HolinkaUnited States Department of Agriculture KBKeith A. BerggrenPrinceton University

Discussion

Member takes

Implication

May expedite ASFV genetic studies; leaves open translation to veterinary applications pending validation.

Key Points

To establish an efficient CRISPR-Cas9 gene editing platform to generate recombinant African swine fever viruses and overcome limitations of traditional homologous recombination.
Applied the CRISPR-Cas9 system in primary swine macrophage cell substrates to target and edit the genome of the virulent field strain ASFV Georgia07.
Targeted the non-essential viral gene 8-DR for deletion to produce recombinant virus constructs.
CRISPR-Cas9 successfully mediated targeted deletion of the non-essential 8-DR gene from the virulent Georgia07 viral genome.
The gene-editing method provided a more robust and efficient alternative to standard homologous recombination for generating and isolating recombinant viruses.

Structured PICO

Population

Swine macrophages infected with highly virulent field strain Georgia07 of African swine fever virus (ASFV)

Intervention

CRISPR-Cas9 gene editing system to delete the non-essential gene 8-DR

Comparator

Traditional homologous recombination

Outcome

Production of recombinant ASFVs

CRISPR-Cas9 provides a robust and efficient system to produce recombinant African swine fever viruses, which may facilitate the development of live-attenuated vaccines.

Limitations

Less than 1% recombinant virus on the initial transfection
CRISPR/Cas9 may be unable to edit all copies of the ASFV genome in a single cell before viral DNA is packaged

Cite This Study

Borca et al. (2018) studied African swine fever virus (ASFV). CRISPR-Cas9 gene editing system vs. Traditional homologous recombination was evaluated on Frequency of recombination (ratio of recombinant to parental viruses). CRISPR-Cas9 increased the efficiency of developing recombinant African swine fever viruses, achieving a recombination frequency of 1 in 122 to 178 compared to <1 in 1,000,000 with traditional methods.

synapsesocial.com/papers/6aa2a5f45d6e77078aa89603 https://doi.org/10.1038/s41598-018-21575-8

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CRISPR-Cas9, a tool to efficiently increase the development of recombinant African swine fever viruses

Discussion

Member takes

Implication

Key Points

Structured PICO

Limitations

Cite This Study

CRISPR-Cas9, a tool to efficiently increase the development of recombinant African swine fever viruses

Discussion

Member takes

Implication

Key Points

Structured PICO

Limitations

Cite This Study

Also Consider

Also Consider